[go: up one dir, main page]

WO2007016372A2 - Methode servant a regenerer un systeme immunitaire - Google Patents

Methode servant a regenerer un systeme immunitaire Download PDF

Info

Publication number
WO2007016372A2
WO2007016372A2 PCT/US2006/029483 US2006029483W WO2007016372A2 WO 2007016372 A2 WO2007016372 A2 WO 2007016372A2 US 2006029483 W US2006029483 W US 2006029483W WO 2007016372 A2 WO2007016372 A2 WO 2007016372A2
Authority
WO
WIPO (PCT)
Prior art keywords
subject
cells
compounds
stem cells
transplanting
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2006/029483
Other languages
English (en)
Other versions
WO2007016372A3 (fr
Inventor
Mitchell J. Ghen
Ramesh Roshan
Romin Roshan
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Eden Biotech Ltd
Original Assignee
Eden Biotech Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Eden Biotech Ltd filed Critical Eden Biotech Ltd
Priority to CA002617108A priority Critical patent/CA2617108A1/fr
Priority to EP06813245A priority patent/EP1919489A4/fr
Priority to US11/997,069 priority patent/US20090202496A1/en
Priority to JP2008524210A priority patent/JP2009502176A/ja
Priority to BRPI0614919-7A priority patent/BRPI0614919A2/pt
Priority to AU2006275625A priority patent/AU2006275625A1/en
Publication of WO2007016372A2 publication Critical patent/WO2007016372A2/fr
Anticipated expiration legal-status Critical
Publication of WO2007016372A3 publication Critical patent/WO2007016372A3/fr
Priority to GB0803704A priority patent/GB2446310A/en
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/26—Lymph; Lymph nodes; Thymus; Spleen; Splenocytes; Thymocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/20—Cytokines; Chemokines
    • C12N2501/23—Interleukins [IL]
    • C12N2501/235—Leukemia inhibitory factor [LIF]

Definitions

  • This invention relates to regenerating an immune system and particularly to treating a subject infected with human immunodeficiency virus (HIV), and an isolated and purified stem cell line for doing so.
  • HAV human immunodeficiency virus
  • HSC allogeneic or autologous hematopoietic stem cells
  • the human immunodeficiency virus primarily infects cells of the immune system, such as T-cells, macrophages, and dendritic cells.
  • the virus infects immune cells via the cell receptor CD4 and a co-receptor which, in primary HIV infection, is the CCR5 receptor. Both receptors are required for the virus to enter the cell where it then replicates.
  • the virus replicates rapidly and infects cells throughout the body, particularly in lymphoid organs.
  • the infection stimulates the immune system to attack the infected cells and reduce HIV levels, but the virus rapidly mutates to avoid this immune attack and so the HIV infection remains.
  • An infected person may then remain in a symptom-free latent stage of infection for years, despite continuous replication of HIV in infected organs.
  • Latency may be broken if the immune system is stimulated, such as during infection by a different pathogen or by activation of CD4+ T-cells in infected lymphoid organs. HIV-infected T-cells are consequently destroyed as large amounts of virus are produced and released from the cells. Under certain conditions, apoptosis may be induced in both infected and uninfected T-cells thus further depleting the T-cell population. As the number of functional T-cells rapidly declines, immune function is compromised, and AIDS (acquired immunodeficiency syndrome) symptoms appear. The course of AIDS is characteristically described as a net change between destruction and production of CD4+ T-cells.
  • HAART nti retroviral therapy
  • HSC transplantation in AIDS patients is of limited use even when combined with HAART.
  • the HSC differentiate into hematopoitic progenitor cells, which give rise to several types of immune cells.
  • Progenitor cells that give rise to T-cells migrate to the thymus, where T-cell differentiation takes place. Mature T-cells
  • I 5 bearing CD4 and CCR5 receptors are then released from the thymus.
  • the health and function of the thymus is critical for the formation of mature T-cells. While the HSC themselves cannot be infected by HIV, these mature T-cells will be readily infected by the virus. AIDS symptoms are temporarily diminished by the HSC transplantation as healthy immune cells are regenerated, but return as the regenerated cells become
  • HSC transplantation is highly dependent on stringent immunosuppressive therapy to prevent immune rejection of the transplanted cells.
  • radiation and/or chemotherapy has been used to destroy the recipient's immune cells prior to transplanting HSC.
  • a person whose immune system is 25 already weakened by HIV infection is subjected to treatments that further deplete immune cells and leave the patient with no defense against subsequent infection.
  • a new method of regenerating an immune system comprises transplanting into the subject a plurality of isolated and purified hematopoeitic stem cells incapable of expressing a functional CCR5 surface receptor, wherein the transplanted cells differentiate into mature immune cells.
  • a nutritional regimen comprising one or more compounds that improve the nutritional status of the subject and one or more anti-microbial compounds is administered to the subject.
  • the nutritional regimen further comprises one or more anti-inflammatory compounds and one or more compounds that stimulate one or both of the growth and the function of the thymus of the subject, and such compounds are administered concurrently with and subsequent to the transplantation of the hematopoetic stem cells.
  • a method of treating a subject infected with HIV comprises transplanting into the subject a plurality of isolated and purified hematopoeitic stem cells incapable of expressing a functional CCR5 surface receptor, wherein the transferred cells differentiate into mature immune cells.
  • the method further comprises transplanting a plurality of mesenchymal stem cells into the subject.
  • the invention is directed to an isolated and purified cell line of hematopoeitic stem cells (HSC) that are incapable of expressing a functional CCR5 receptor on the cell surface ("the CCR5 ⁇ 32 cells").
  • HSC hematopoeitic stem cells
  • the invention is further directed to a method of regenerating the immune system in a subject in need thereof by transplanting CCR5 ⁇ 32 into the recipient subject. Because mature immune cells derived from CCR5 ⁇ 32 cells cannot express functional CCR5 receptors, they will be resistant to infection by HIV and other pathogens that use the CCR5 receptor to infect cells. (Agrawal et a/., J.
  • the method of regenerating the immune system also includes administration of a nutritional regimen to the patient that optimizes conditions for CCR5 ⁇ 32 cell transplantation.
  • CCR5 receptor - Cell surface protein utilized by HIV to infect host cells often referred to as a coreceptor because HIV also requires CD4 receptors to infect a host cell.
  • CD4 receptor - Cell surface protein utilized by HIV to infect host cells.
  • a cell Express, expression - Production of a protein, i.e., a cell will "express” a protein when it synthesizes that protein; a protein, such as a receptor, is “expressed” when it is synthesized by a cell; and “receptor expression” is the synthesis of a receptor protein.
  • Graft v. host response The destruction of host cells by donor cells transferred into the host.
  • Host v. graft response The destruction of donor cells transferred into a host by the host immune cells.
  • Immune cell A cell capable of an immune response
  • Immune response A response made by the immune system to a foreign substance, includes transplant rejection, antibody production, inflammation, and response of antigen specific lymphocytes to antigen.
  • Immune system The bodily system that protects the body from foreign substances, cells, and tissues by producing the immune response and that includes especially the thymus, spleen, lymph nodes, special deposits of lymphoid tissue (as in the gastrointestinal tract and bone marrow), lymphocytes including the B-cells and T-cells, and antibodies.
  • Immunosuppression - Prevention, diminution, or delay of an immune response Mature immune cell - cell of the immune system that can not differentiate into another cell type.
  • MSC mesenchymal stem cell - Human bone marrow stromal stem cells that are pluripotent progenitor cells with the ability to generate cartilage, bone, muscle, tendon, ligament and fat tissues.
  • Nutraceutical A food or naturally occurring food supplement thought to have a beneficial effect on human health.
  • the CCR5 ⁇ 32 Cell Line The HSC cell line, CCR5 ⁇ 32, is a human hematopoeitic stem cell line derived from a human fetus that bears a 32-base pair deletion on chromosome 3 in the coding region of the CCR5 gene. The deletion is described in McNicholl, et al., Host genes and HIV: the role of the chemokine receptor gene CCR5 and its allele ( ⁇ 32 CCR5), Emerging Infectious Diseases 3: 261-271, 1997, and in U. S. Patent No. 6,692,938, both incorporated herein by reference.
  • the CCR5 ⁇ 32 cells differentiate into functional immune cells that lack CCR5 receptors. Because the cells cannot produce the CCR5 receptor, pathogens, such as HIV and small pox virus, that require CCR5 receptor to infect cells cannot invade these cells and replicate within them. As a result, after CCR5 ⁇ 32 HSC are transplanted into, for example, an HIV infected/AIDS patient, T-cells that differentiate from the CCR5 ⁇ 32 HSC will be resistant to HIV infection. Over time, infected T-cells of the recipient will be replaced by resistant T-cells derived from CCR5 ⁇ 32 HSC, thereby reducing HIV infection and viral replication.
  • pathogens such as HIV and small pox virus
  • the resistant immune cells will also help to repair and regenerate tissues and organs, such as the thymus, bone marrow, and lymphoid tissues, which have been damaged as a result of the HIV infection, thereby providing continued sources of healthy immune cells.
  • tissues and organs such as the thymus, bone marrow, and lymphoid tissues, which have been damaged as a result of the HIV infection, thereby providing continued sources of healthy immune cells.
  • the regenerated immune system is derived from the donor cells, subsequent transplantations from the same source of donor HSC will not be rejected by the regenerated immune system.
  • a specialized nutritional regimen is administered to the recipient before, during, and after HSC transplantation to optimize conditions that help establish the CCR5 ⁇ 32 HSC in the recipient, and to lessen the trauma associated with immunosuppression.
  • the nutritional regimen may additionally reduce viral load.
  • the steps of the nutritional regimen are (1) administration of nutrients to optimize the nutritional status of the recipient, so that the transplanted cells will have a healthy environment in which to become established; (2) administration of anti-microbial compounds to remove and prevent infection in the recipient; (3) administration of neutraceuticals that reduce inflammation in the recipient to enhance the establishment of the transplanted HSC in the recipient; and (4) administration of compounds that stimulate the function and size of the thymus to allow continued propagation and differentiation of the transplanted HSC.
  • the nutritional regimen has three parts: the pre-transplantation phase, the days of HSC transplantation, and the post-transplantation phase.
  • Nutrient compounds administered for step (1) may include antioxidants, such as acetyl-1-carnitine; alpha- lipoic acid; enzymes, such as Coenzyme QlO; vitamins, such as Vitamins C and E; thyroid extract; and nutraceuticals, such as undenatured whey protein, blueberry extract and resveratrol.
  • Anti-microbial compounds administered for step (2) include natural antimicrobial agents, such as allicin, oregabiotic, colloidal silver, oil of oregano, Artemesia with citrus seed extract; and ozone.
  • the nutritional regimen includes all four steps and is administered at selected intervals of approximately every 30 days for at least 4 months for up to one year after the transplantation procedure.
  • the compounds listed above for steps (1) and (2) are again administered and are supplemented with compounds that stimulate the thymus and suppress inflammation (steps 3 and 4).
  • Compounds that stimulate growth and function of the thymus include fatty acids, such as alpha and gamma linolenic acid, eiocosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and linoleic acid; phosphatidyl choline; glutathione; carotenoids; methyl cobalamine; thyroid stimulating agents, such as thyroid hormone and thyroid extracts; Vitamins A and D; minerals, such as zinc, calcium, magnesium, potassium, chromium, selenium, germanium, rubidium; human growth hormone; amino acids; adenosine monophosphate; and alkylglycerols such as shark liver oil.
  • fatty acids such as alpha and gamma linolenic acid, eiocosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and linoleic acid
  • Anti-inflammatory compounds include butryn, cinnamon bark, curcumin, kaprex, RPR, quercitin, essential fatty acids, and vitamins C and E.
  • Therapeutic amounts of each of the compounds given are known to those of ordinary skill in the art, and, for each stage, the amount, specific compound, and method and timing of administration of each compound may be optimized for the recipient.
  • Pre-transplantation phase and transplantation phase are known to those of ordinary skill in the art, and, for each stage, the amount, specific compound, and method and timing of administration of each compound may be optimized for the recipient.
  • the pre-transplantation phase begins two days before the first transplantation procedure.
  • the same regimen is also administered on the day(s) of HSC transplantation.
  • the following compounds are administered daily to the recipient via intravenous infusion:
  • Vitamin C 50,000 mg per day methocarbamol, 750 mg per day
  • Vitamin E 1000 units per day
  • Coenzyme QlO 400 mg per day acetyl-1-camitine, 3 g per day alpha-lipoic acid, 1000 mg per day ozone, 20 ml three times per day mild silver protein or colloidal silver, 5-1OmI administered in 5% Dextrose in water
  • the following compounds are administered orally each day: allicin, 450 mg, 3-4 times per day artemesia, 150 mg per day oregabiotic, 500 mg, 1-4 times per day resveratol, 50-100 mg per day ARMOUR ® thyroid, 0.5 to 1 grain (Forest Pharmaceuticals, Inc.) that is greater than 2 mu/ml.
  • each of the following compounds is administered at 20, 29, 61, 90, 118, and 365 days after the transplantation to stimulate the thymus.
  • Vitamin C 50,000 mg, intravenously
  • Super Immune 500 ml in water intravenously. Includes 25,000 mg of vitamin C, 200 mg of B6, 1 cc of B-complex, 10 cc of 10% calcium gluconate, 2000 mg magnesium sulfate, 750 mg pantothenic acid, 15 mg folic acid, 400 ⁇ g selenium, 2 cc of adenosine monophosphate or 7 cc of Glycyrrhizin, 10 cc of glutathione, 10 cc of 50 mg/ml taurine, 2 cc of either hydroxycobalamine or methylcobalamine, 2 cc of multi-trace minerals, and 5 cc of zinc.
  • Vitamin A 50,000 i.u., intramuscularly mixed carotenoids, 200,000 i.u., orally s zinc, 150 mg, orally phosphatidyl choline, 500 mg in tandom slow intravenous push with 1500 mg glutathione human growth hormone, 8 units, subcutaneously adenosine monophosphate, 40 mg, intravenously o dipyridamole, 50 mg, 2-3 times/day, orally
  • N-acetyl cysteine 8000mg/day, orally butryn, 3 times/day, orally
  • L-glutamine 5 g, 3 times/day, orally s L-arginine, 6 g, orally
  • Vitamin D3 10,000 i.u., intramuscularly
  • ARMOUR ® thyroid thyroid extract (thyroid extract) (Forest Pharmaceuticals, Inc.), 1 grain at greater than 2mu/ml concentration, orally 0 methylcobalamine, 5 mg, 4 times/day, orally folic acid, 15 mg, orally
  • Vitamin E 1000 i.u., orally alkylglycerols, (shark liver oil) 400mg on days 90, 118, and 365, orally quercitin, 3 times/day, orally S KaprexTM (Metagenics), 1 tablet 3 times/day, orally
  • Bone marrow transplantation recipients are generally subjected to toxic methods of immunosuppression such as chemotherapy and radiation to destroy 5 the recipient's own hematopoietic cells. These procedures can give rise to anemia, neutropenia, and increased susceptibility to infection. Infection subsequent to severe immunosuppression can result in death of the recipient.
  • T-cells are tolerant of both donor and recipient antigens on immune cells, the recipient subject's immune cells will not destroy the donor cells, and subsequent cell transplantations from the same donor source will not require further immunosuppression.
  • Purified hematopoietic stem cell grafts induce tolerance to alloantigens and can mediate positive and negative T cell selection.
  • immunosuppression may be accomplished by any method known to those in the art, less traumatic procedures are preferred to encourage mixed chimerism and to minimize trauma to the transplantation recipient.
  • One preferred method of immunosuppression that also supports growth and development of the transplanted HSC is short-term treatment with mycophenolate mofetil in combination with administration of Vitamin D and co-transplantation of HSC with mesechymal stem cells (MSC).
  • mycophenolate mofetil (MMF) is administered to the recipient intravenously at a dose of 1-3 g over a period of 2-4 hours prior to transplantation on each day that an HSC transplantation is performed. Following the final transplantation procedure, a 1-3 g/day dose of MMF is administered to the recipient orally for 13 days.
  • Vitamin D is also administered at 50,000 IU given orally on each day that MMF is administered.
  • MSC have immunosuppressive activity and appear to enhance the establishment of donor HSC in recipients.
  • K. Le Blanc, et al. Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. Scandinavian J. Immunol. 57: 11-20, 2003; in A. Bacigalupo, Mesenchymal stem cells and hematopoietic stem cell transplantation. Best Practice & Research Clinical Haematology 17: 387-399, 2004; W. E. Fibbe and W.A. Noort, Mesenchymal stem cells and hematopoietic stem cell transplantation. Ann. N.Y. Acad.
  • MSC may be isolated from adult human bone marrow, propagated in culture, and prepared for transplantation as described, for example, in K. Le Blanc, et al., Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. Scandinavian J. Immunol. 57: 11-20, 2003; O.N. Ko ⁇ , et a/., Allogeneic mesenchymal stem cell infusion for treatment of metachromatic leukodystrophy (MLD) and Hurler syndrome (MPSOIH).
  • MLD metachromatic leukodystrophy
  • MPSOIH Hurler syndrome
  • kits for immunosuppression include, but are not limited to, radiation, administration of chemotherapeutic agents such as cyclosporine, busulphan, cyclophosphamide, methotrexate, and administration of alemtuzumab or other antibodies.
  • chemotherapeutic agents such as cyclosporine, busulphan, cyclophosphamide, methotrexate, and administration of alemtuzumab or other antibodies.
  • methods of the present invention are accomplished in a way which reduces or eliminates the need for any radiation to be used on the subject.
  • the CCR5 ⁇ 32 HSC may be transferred by any appropriate methods known in the art, such as peripherally; by intra-bone marrow injection ⁇ e.g., Castello et al., Intra- bone marrow injection of bone marrow and cord blood cells: An alternative way of transplantation associated with a higher seeding efficiency. Experimental Hematology 32: 782-787, 2004; by any standard method of intra-thecal injection,, by intrathymic injection (e.g.Trani et al., CD25 + immunoregulatory CD4 T cells mediate acquired central transplantation tolerance. J. Immunology 170: 279-286, 2003); by direct organ injection, or by other appropriate means.
  • intra-bone marrow injection e.g., Castello et al., Intra- bone marrow injection of bone marrow and cord blood cells: An alternative way of transplantation associated with a higher seeding efficiency.
  • Experimental Hematology 32: 782-787, 2004 by any standard method of intra-thecal injection, by intra
  • Combinations of different methods may be used for successive transplantations to increase the probability that the HSC will become established in the recipient.
  • Serial transplantations of CCR5 ⁇ 32 HSC will be performed to increase the likelihood that the transplanted cells will become established and regenerate the recipient's immune system. Immunosuppression will be required for the first serial transplantations of CCR5 ⁇ 32 HSC to promote chimerism, but may not be required for subsequent transplantations of CCR5 ⁇ 32 HSC.
  • CCR5 ⁇ 32 HSC will be transplanted each day for three sequential days. Different methods of transplantation may be employed on each day of transplantation. For example, IBMI and peripheral transplantations may be performed on the same day, or IBMI may be used for the first transplantation, and intrathymic injection or peripheral transplantation may be used for the second and third transplantations.
  • transplantations of CCR5 ⁇ 32 HSC may again be made one year later if necessary.
  • the number of transplantations and mode of administration of the CCR5 ⁇ 32 HSC may be optimized for each recipient.
  • the transplantation of CCR5 ⁇ 32 HSC may be made each day for three successive days. At least one of the three transplantations of CCR5 ⁇ 32 HSC is into bone marrow.
  • the recipient is anesthetized using both local infiltration and light general anesthesia/analgesia procedures.
  • a bone marrow cannula is utilized to enter the bone marrow through the sternum or other bone marrow site.
  • Approximately 4 x 10 5 to 1 x 10 5 CCR5 ⁇ 32 HSC in ImI of a solution of 95% PBS + 5% DMSO are injected into the bone marrow.
  • Alternative routes of CCR5 ⁇ 32 HSC transplantation are injected into the bone marrow.
  • thymic injection may be used as an alternative method of transplantation for CCR5 ⁇ 32 HSC.
  • the recipient is prepared as described for IBMI. Ultrasound is used to guide a needle into the thymus, and approximately 1-10 x 10 6 CCR5 ⁇ 32 HSC are injected directly into the thymus. During this surgical procedure, 2- 10 x 10 6 MSC in ImI of a solution of 95% PBS + 5% DMSO are transplanted intravenously into the recipient.
  • CCR5 ⁇ 32 HSC may also be transplanted peripherally according to known methods. For this procedure, 7-9 x 10 6 CCR5 ⁇ 32 HSC cells are administered intravenously. Co-transplantation of MSC
  • MSC are transplanted at the same time as the CCR5 ⁇ 32 HSC cells.
  • 3 x 10 6 MSC are suspended in 100 ml of 0.9% normal saline and transplanted intravenously into the recipient.
  • SNP analysis may be used to monitor the donor: recipient blood cell ratio following the HSC transplantation to verify the formation of chimerism, i.e., that both donor and recipient immune cells are present.
  • SNP analysis may be performed according to Harries, L, et al., Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction. Bone Marrow Transplantation 35: 283-290, 2005. Differentiation of donor HSC may be monitored by flow cytometry targeting CD8, CD4 and CD45RA receptors. A relative increase in CD4 and CD45RA receptors indicates HSC differentiation.
  • Thymic T-cell output may be measured by TREC (T cell Receptor excision circles) quantification as described, for example, in Weinberg, K. et al., Factors affecting thymic function after allogeneic hematopoietic stem cell transplantation. Transplantation 97: 1458-1466, 2001.
  • TREC T cell Receptor excision circles
  • CCR5-deficient mice do not develop tuberculosis when infected by Mycobacterium tuberculosis, but instead mount a protective immune response to the infection which prevents development of the disease. (Algood, H. M. S. and Flynn, J. L., CCR5-deficient mice control Mycobacterium tuberculosis infection despite increased pulmonary lymphocytic infiltration. J. Immunol. 173: 3287-3296. 2004, incorporated herein by reference).
  • CCR5 ⁇ 32 mutation is thought to have evolved as a protective mechanism against smallpox. (Galvani, A. P. and Slatkin, M., Evaluating plague and smallpox as historical selective pressures for the CCR5 ⁇ 32 HIV-resistance allele. P.N.A.S. USA 100: 15276-15279, 2003, incorporated herein by reference).
  • the cell line can be isolated, purified, and expanded from fetal liver tissue that is homozygous for the 32-base pair deletion in the CCR5 receptor gene.
  • fetal liver tissue was pressed through a wire mesh sieve to separate the cells, which were then placed in culture.
  • HSC were purified from liver cells by culturing and expanding in selective medium over two to three passages, using standard tissue culture techniques.
  • CCR5 ⁇ 32 medium comprised Dulbecco's modified Eagle's medium IX with glucose and L-glutamine, 29.4% fetal bovine serum (FBS), penicillin/streptomycin, lymphocyte inhibiting factor (LIF) 20 ng/ml, basic fibroblast growth factor (FGF) 100 ng/ml,and stem cell factor (SCF) 4 ng/ml.
  • the medium is supplemented with 4.2% non-essential amino acids, L- glutamine 2.34 mg/rril, 2-mercaptoethanol 2.22 mg/ml, and sodium pyruvate 0.22 mg/ml.
  • the pH of the medium was substantially maintained at 7.4.
  • CCR5 ⁇ 32 medium was also used for propagation of the CCR5 ⁇ 32 cells.
  • the cells were plated at 1.8xlO 5 cells/cm 2 in a 75 cm 2 flask with a 0.2 micrometer vent cap. The cultures were maintained at 37°C in a humidified incubator containing 95% air and 5% CO 2 and were subcultured prior to confluence.
  • Cells were extracted from the medium by dialysis for use or for storage.
  • the media containing the cells was placed in MWCO 250,000 dialysis tubing and dialysed against a solution of phosphate buffered saline (PBS).
  • PBS phosphate buffered saline
  • the cells are frozen in 95% PBS/5% DMSO and stored in liquid nitrogen.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Virology (AREA)
  • Hematology (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Reproductive Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Molecular Biology (AREA)
  • AIDS & HIV (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

On utilise une lignée cellulaire isolée et purifiée de cellules souches hématopoïétiques (HSC) incapables d'exprimer le récepteur CCR5 sur leur surface, ('cellules CCR5Δ32'), afin de régénérer le système immunitaire d'un individu et, en particulier, de traiter un sujet infecté par le virus de l'immunodéficience humaine (VIH). On met en application cette méthode par transplantation de CCR5Δ32 dans l'individu récepteur. Etant donné que les cellules immunes matures dérivées des cellules CCR5Δ32 ne peuvent pas exprimer les récepteurs fonctionnels CCR5, elles seront résistantes à l'infection par VIH ou d'autres pathogènes utilisant ce récepteur CCR5 pour infecter des cellules. Un mode de réalisation de l'invention consiste à administrer un régime nutritionnel au patient, ce qui permet d'optimiser les conditions de transplantation des cellules CCR5Δ32. Un autre mode de réalisation consiste à co-transplanter des cellules mésenchymateuses avec HSC, de façon à augmenter la croissance et le développement de HSC transplanté.
PCT/US2006/029483 2005-07-28 2006-07-27 Methode servant a regenerer un systeme immunitaire Ceased WO2007016372A2 (fr)

Priority Applications (7)

Application Number Priority Date Filing Date Title
CA002617108A CA2617108A1 (fr) 2005-07-28 2006-07-27 Methode servant a regenerer un systeme immunitaire
EP06813245A EP1919489A4 (fr) 2005-07-28 2006-07-27 Methode servant a regenerer un systeme immunitaire
US11/997,069 US20090202496A1 (en) 2005-07-28 2006-07-27 Method for regenerating an immune system
JP2008524210A JP2009502176A (ja) 2005-07-28 2006-07-27 免疫系の再生方法
BRPI0614919-7A BRPI0614919A2 (pt) 2005-07-28 2006-07-27 método para regenerar um sistema imune
AU2006275625A AU2006275625A1 (en) 2005-07-28 2006-07-27 Method for regenerating an immune system
GB0803704A GB2446310A (en) 2005-07-28 2008-02-28 Method for regenerating an immune system

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US70307305P 2005-07-28 2005-07-28
US60/703,073 2005-07-28

Publications (2)

Publication Number Publication Date
WO2007016372A2 true WO2007016372A2 (fr) 2007-02-08
WO2007016372A3 WO2007016372A3 (fr) 2008-01-31

Family

ID=37709223

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2006/029483 Ceased WO2007016372A2 (fr) 2005-07-28 2006-07-27 Methode servant a regenerer un systeme immunitaire

Country Status (9)

Country Link
US (1) US20090202496A1 (fr)
EP (1) EP1919489A4 (fr)
JP (1) JP2009502176A (fr)
KR (1) KR20080048463A (fr)
AU (1) AU2006275625A1 (fr)
BR (1) BRPI0614919A2 (fr)
CA (1) CA2617108A1 (fr)
GB (1) GB2446310A (fr)
WO (1) WO2007016372A2 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2041269A4 (fr) * 2006-06-28 2009-11-25 Searete Llc Procédés destinés à modifier la sensibilité cellulaire à une infection
US10918672B1 (en) 2016-04-07 2021-02-16 The Administrators Of The Tulane Educational Fund Small tissue CCR5−MSCs for treatment of HIV
US20240095677A1 (en) * 2019-07-11 2024-03-21 David Mroczka Method and system for guidance of artificial intelligence and human agent teaming

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2012275526A1 (en) * 2011-06-29 2014-01-16 Ronilu Development Corporation Prevention and treatment of HIV infection
WO2014151994A1 (fr) 2013-03-15 2014-09-25 Kambiz Shekdar Édition de génome a l'aide d'oligonucléotides effecteurs pour un traitement thérapeutique
CN111996164A (zh) * 2020-09-10 2020-11-27 聊城市人民医院 一种间充质干细胞的无血清抗衰老培养基

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4117904B2 (ja) * 1996-03-01 2008-07-16 エーロスクレーン エッス．アー． Ｃ−ｃ ｃｋｒ−５，ｃｃ−ケモカインレセプタ、その誘導体及びこれらの利用
US6153431A (en) * 1997-05-30 2000-11-28 Fond Mondiale Rech & Prev Sida Human immunodeficiency virus co-receptor variants associated with resistance to virus infection
US20030017587A1 (en) * 2001-07-18 2003-01-23 Rader William C. Embryonic stem cells, clinical applications and methods for expanding in vitro
US6877574B2 (en) * 2001-09-14 2005-04-12 Massachusetts Institute Of Technology Modular robotic teaching tool
US20030099621A1 (en) * 2001-11-29 2003-05-29 Robert Chow Stem cell screening and transplantation therapy for HIV infection
WO2004013330A1 (fr) * 2002-07-26 2004-02-12 Consejo Superior De Investigaciones Científicas Constructions genetiques multifonctionnelles a capacite d'inhibition elevee de l'expression du ccr5 dans la surface cellulaire

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of EP1919489A4 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2041269A4 (fr) * 2006-06-28 2009-11-25 Searete Llc Procédés destinés à modifier la sensibilité cellulaire à une infection
US10918672B1 (en) 2016-04-07 2021-02-16 The Administrators Of The Tulane Educational Fund Small tissue CCR5−MSCs for treatment of HIV
US20240095677A1 (en) * 2019-07-11 2024-03-21 David Mroczka Method and system for guidance of artificial intelligence and human agent teaming

Also Published As

Publication number Publication date
BRPI0614919A2 (pt) 2011-04-19
US20090202496A1 (en) 2009-08-13
AU2006275625A2 (en) 2008-05-01
JP2009502176A (ja) 2009-01-29
CA2617108A1 (fr) 2007-02-08
EP1919489A4 (fr) 2009-05-06
WO2007016372A3 (fr) 2008-01-31
GB2446310A (en) 2008-08-06
GB0803704D0 (en) 2008-04-09
AU2006275625A1 (en) 2007-02-08
KR20080048463A (ko) 2008-06-02
EP1919489A2 (fr) 2008-05-14

Similar Documents

Publication Publication Date Title
JP6449199B2 (ja) ヒト促進細胞およびその使用
RU2497530C2 (ru) Способ лечения иммунной дисфункции, такой как реакция "трансплантат против хозяина" или "хозяин против трансплантата"
US8075921B2 (en) Rapamycin-resistant T cells and therapeutic uses thereof
US10792309B2 (en) Cell therapy composition for preventing or treating immune disease comprising mesenchymal stem cells and immunoregulatory t-cells as active ingredient
RU2493869C2 (ru) Tr1-КЛЕТКИ, МЕЗЕНХИМАЛЬНЫЕ СТВОЛОВЫЕ КЛЕТКИ И ИХ ПРИМЕНЕНИЕ
US20150104471A1 (en) Universal donor-derived tolerogenic cells for inducing non-syngeneic transplantation tolerance
EP2613801B1 (fr) LYMPHOCYTES T À MÉMOIRE CENTRALE ANTI-TIERS, ET UTILISATION DE CEUX-CI DANS UN TRAITEMENT & xA;'ANTI-LEUKEMIA/LYMPHOMA'
US20120141434A1 (en) Methods for Selecting Expanded Stem Cell Populations
Francese et al. Immunological and regenerative properties of cord blood stem cells
Lu et al. Regulatory T cells induce transplant immune tolerance
JP2009536163A (ja) 免疫学的寛容および調節性前駆細胞
US10751368B2 (en) Methods of transplantation and disease treatment
US20090202496A1 (en) Method for regenerating an immune system
US7544355B2 (en) Methods and compositions for allogeneic transplantation
CA2529244C (fr) Lymphocytes t resistant a la rapamycine et leurs utilisations therapeutiques
Touraine Transplantation of human fetal liver cells into children or human fetuses
US20240108655A1 (en) Methods of producing mixed chimerism after a solid organ transplant
Schatton et al. IN VIVO IMMUNOMODULATORY FUNCTION OF ABCB5+ DERMAL MESENCHYMAL STEM CELLS.
HK1163738A (en) Methods for selecting expanded stem cell populations
IL189688A (en) Universal donor cells provide tolerance for tolerance to a non-self-donor

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application
ENP Entry into the national phase

Ref document number: 2617108

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2008524210

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 1020087004442

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 2006275625

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 0803704

Country of ref document: GB

Kind code of ref document: A

Free format text: PCT FILING DATE = 20060727

WWE Wipo information: entry into national phase

Ref document number: 0803704.6

Country of ref document: GB

Ref document number: 2006813245

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2006275625

Country of ref document: AU

Date of ref document: 20060727

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 11997069

Country of ref document: US

ENP Entry into the national phase

Ref document number: PI0614919

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20080128